miR-410调节NF-κB信号通路影响脂多糖刺激的牙髓干细胞增殖及炎症反应

梁敏; 杨文; 文涛

doi:10.13898/j.cnki.issn.2097-5252.2025.07.003

miR-410调节NF-κB信号通路影响脂多糖刺激的牙髓干细胞增殖及炎症反应

Effects of miR-410 regulating NF-κB signaling pathway on lipopolysaccharide-stimulated proliferation and inflammatory response of dental pulp stem cells

摘要

摘要:
目的探究miR-410调节NF-κB信号通路对脂多糖(LPS)刺激的牙髓干细胞增殖能力及炎症因子分泌的影响。
方法采用qRT-PCR检测LPS刺激不同时间(0、12、24、48 h)牙髓干细胞中miR-410表达变化。将牙髓干细胞分为对照(Con)组、LPS组(10 ng/mL LPS处理48 h)、LPS+miR-NC组(转染miR-NC后经LPS处理48 h)、LPS+miR-410 mimics组(转染miR-410后经LPS处理48 h)、LPS+miR-410 mimics+PMA组(转染miR-410后经PMA和LPS处理48 h)。通过qRT-PCR检测miR-410表达，MTT实验检测细胞增殖能力，ELISA检测炎症因子白细胞介素(IL)-6、IL-1β和肿瘤坏死因子α(TNF-α)水平，蛋白免疫印迹法检测细胞周期相关蛋白CyclinD1及NF-κB信号通路相关蛋白p-p65 NF-κB、p-IκBα表达。
结果随着LPS处理时间延长，牙髓干细胞中miR-410表达水平逐渐下降(P < 0.01)。与Con组相比，LPS组牙髓干细胞中miR-410表达水平、细胞存活率、CyclinD1蛋白表达水平均降低，IL-6、IL-1β、TNF-α水平均升高，p-p65 NF-κB、p-IκBα蛋白表达水平均升高(P < 0.05)。与LPS+miR-NC组相比，LPS+miR-410 mimics组牙髓干细胞中miR-410表达水平、细胞存活率、CyclinD1蛋白表达水平均增加，IL-6、IL-1β、TNF-α水平均降低，p-p65 NF-κB、p-IκBα蛋白表达水平均降低(P < 0.05)。与LPS+miR-410 mimics组相比，LPS+miR-410 mimics+PMA组牙髓干细胞存活率、CyclinD1蛋白表达水平均明显降低，IL-6、IL-1β、TNF-α水平均明显升高(P < 0.01)。
结论过表达miR-410可通过抑制NF-κB信号通路促进LPS刺激的牙髓干细胞增殖能力，抑制炎症因子分泌。

Abstract:
Objective To investigate the effect of miR-410 on the lipopolysaccharide (LPS)-stimulated proliferation ability and inflammatory factor secretion of dental pulp stem cell by regulating NF-κB signaling pathway.
Methods The expression changes of miR-410 in dental pulp stem cells stimulated with LPS at different times (0, 12, 24, 48 h) were detected by qRT-PCR.Dental pulp stem cells were divided into control (Con) group, LPS group (treated with 10 ng/mL LPS for 48 h), LPS+miR-NC group (transfected with miR-NC and then treated with LPS for 48 h), LPS+miR-410 mimics group (transfected with miR-410 and then treated with LPS for 48 h), LPS+miR-410 mimics+PMA group (transfected with miR-410 and then treated with PMA and LPS for 48 h).The expression of miR-410 was detected by qRT-PCR, the cell proliferation was detected by MTT assay, the secretion of inflammatory factors interleukin (IL)-6, IL-1β, and tumor necrosis factor α (TNF-α) was detected by ELISA, and the expressions of cell cycle-related protein CyclinD1 and NF-κB signaling pathway-related proteins p-p65 NF-κB and p-IκBα were detected by Western blotting.
Results The expression of miR-410 in dental pulp stem cells was gradually decreased with the increase of LPS treatment time (P < 0.01).Compared with the Con group, the expression levels of miR-410, cell survival rate, and expression levels CyclinD1 protein of dental pulp stem cells in the LPS group were decreased, while the levels of IL-6, IL-1β, and TNF-α were increased, the protein expression levels of p-p65 NF-κB and p-IκBα were increased (P < 0.05).Compared with the LPS+miR-NC group, the expression level of miR-410, cell survival rate, and CyclinD1 protein expression level of dental pulp stem cells in the LPS+miR-410 mimics group were increased, while the expression levels of p-p65 NF-κB and p-IκBα were decreased (P < 0.05).Compared with the LPS+miR-410 mimics group, the cell survival rate and CyclinD1 protein expression level of dental pulp stem cells in the LPS+miR-410 mimics+PMA group were significantly decreased, while the levels of IL-6, IL-1β, and TNF-α were significantly increased (P < 0.01).
Conclusions Overexpression of miR-410 can promote the proliferation ability of LPS-stimulated dental pulp stem cells and inhibit the secretion of inflammatory factors by inhibiting the NF-κB signaling pathway.

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